Objective: The present study aims to elucidate the significance of immune cell infiltration in Coronavirus disease 2019 (COVID-19) myocarditis and identify potential diagnostic markers for this condition. Myocarditis, an inflammatory cardiac disease, primarily results from viral infections. Although the association between COVID-19 and myocarditis is well-established, the specific mechanism(s) underlying this relationship remain incompletely understood. Methods: The GSE53607 and GSE35182 datasets were obtained from the GEO database, which contains samples from a mouse model for viral myocarditis. Differentially expressed genes (DEGs) and candidate biomarkers were selected using the LASSO regression model and support vector machine recursive feature elimination (SVM-RFE) analysis. Subsequently, the diagnostic potential of these biomarkers was evaluated by calculating the area under the receiver operating characteristic curve (AUC). Further validation of the biomarkers was conducted using the GSE183850 dataset, which consists of samples from patients with COVID-19 myocarditis. In addition, CIBERSORT analysis was employed to estimate the compositional patterns of 22 types of immune cell fractions in merged cohorts. Results: Thirty genes were identified, with a significant proportion of the DEGs being associated with carbohydrate binding, endopeptidase activity, and pathogenic organisms such as Staphylococcus aureus and coronavirus disease. Importantly, gene sets related to the IL6-JAK-STAT3 signaling pathways, inflammatory response, and interferon response exhibited differential activation in viral myocarditis compared to the control group. In addition, in the context of COVID-19 myocarditis patients from the GSE183850 dataset, B2M and C3 were established as diagnostic markers that were subsequently validated (AUC = 0.978 and AUC = 0.956, respectively). Furthermore, analysis of immune cell infiltration revealed correlations between B2M and C3 expression levels and the activation of NK cells, dendritic cells, T cells CD4 memory resting, as well as eosinophils. Conclusion: B2M and C3 have been identified as potential biomarkers for viral myocarditis, providing valuable insights for future investigations into the pathogenesis of COVID-19-associated myocarditis.

Dingji Fumai Decoction (DFD), a traditional herbal concoction, is commonly utilized in therapeutic practice to treat ventricular arrhythmia. However, research into the bioactive components and underlying processes of DFD in Long QT syndrome (LQTS) remains limited. All DFD compounds were gathered from the TCMSP, ETCM, and HERB databases, and the targets of active compounds were investigated using SwissTargetPrediction. The LQTS targets were obtained/screened from the DisGeNET, OMIM, and Malacard databases. The herb-compound-target-disease (H-C-T-D) and PPI networks were built using STRING and analyzed with CytoNCA based on the data obtained earlier. Meanwhile, VarElect was used to determine the relationship between targets and illnesses. The R program is used to enrich Gene Ontology (GO) terminology as well as the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The binding ability of DFD and hub genes was examined using molecular docking, followed by experimental validation of the key findings. A total of 664 DFD and 240 LQTS targets were gathered, with 21 common targets found. The H-C-T-D network demonstrated the links between DFD, active molecules, targets, and LQTS. The PPI network revealed the major targets: KCNH2, HSP90AA1, SCN5A, and CACNA2D1. Further systematic investigation revealed DFD's potential mechanism for protecting against LQTS. Furthermore, molecular docking revealed the interactions of active drugs and targets. Finally, we discovered that DFD enhanced the levels of KCNH2, HSP90AA1, and CACNA2D1 while decreasing the level of SCN5A. DFD has the potential to cure LQTS through a complex method that involves interactions between active chemicals and targets. This study enhances our understanding of the molecular mechanisms underlying DFD's effects on LQTS.

The challenge posed by opioid overdose has become a significant concern for health systems due to the complexities associated with drug prohibition, widespread clinical use, and potential abuse. In response, healthcare professionals have primarily concentrated on mitigating the hallucinogenic and respiratory depressant consequences of opioid overdose to minimize associated risks. However, it is crucial to acknowledge that most opioids possess the capacity to prolong the QT interval, particularly in cases of overdose, thereby potentially resulting in severe ventricular arrhythmias and even sudden death if timely intervention is not implemented. Consequently, alongside addressing the typical adverse effects of opioids, it is imperative to consider their cardiotoxicity. To enhance comprehension of the correlation between opioids and arrhythmias, identify potential targets for prompt intervention, and mitigate the hazards associated with clinical utilization, an exploration of the interaction between drugs and ion channels, as well as their underlying mechanisms, becomes indispensable. This review primarily concentrates on elucidating the impact of opioid drugs on diverse ion channels, investigating recent advancements in this domain, and attaining a deeper understanding of the mechanisms underlying the prolongation of the QT interval by opioid drugs, along with potential interventions.

The potassium channel protein KCNH2 is encoded by KCNH2 gene, and there are more than 300 mutations of KCNH2. Unfolded protein response (UPR) is typically initiated in response to an accumulation of unfolded and/or misfolded proteins in the endoplasmic reticulum (ER). The present study aimed to explore the UPR process and the role of activating transcription factor 6 (ATF6) in the abnormal expression of potassium voltage-gated channel subfamily H member 2 (KCNH2)A561V. The wild-type (wt) KCNH2 and A561V mutant KCNH2 was constructed with his-tag. The 293 cells were used and divided into KCNH2wt+KCNH2A561V, KCNH2wt and KCNH2A561V groups. The expression levels of ATF6 and KCNH2 in different groups were detected by Western blotting, reverse transcription-quantitative PCR, immunofluorescence and immuno-coprecipitation assays. The protein types and abundance of immuno-coprecipitation samples were analyzed by mass spectrometry. The proteomic analysis of the mass spectrometry results was carried out by using the reactome database and GO (Gene Ontology) tool. The mRNA expression levels of KCNH2 and ATF6 in the KCNH2wt+KCNH2A561V group were higher compared with the KCNH2A561V group. However, the full-length protein expression of ATF6 was inhibited, indicating that ATF6 was highly activated and a substantial number of ATF6 was sheared in KCNH2wt+KCNH2A561V group compared with control group. Furthermore, A561V-KCNH2 mutation leading to the accumulation of the immature form of KCNH2 (135 kDa bands) in ER, resulting in the reduction of the ratio of 155 kDa/135 kDa. In addition, the abundance of UPR-related proteins in the KCNH2A561V group was higher compared with the KCNH2wt+KCNH2A561V group. The 'cysteine biosynthetic activity' of GO:0019344 process and the 'positive regulation of cytoplasmic translation activity' of GO:2000767 process in the KCNH2A561V group were higher compared with the KCNH2wt+KCNH2A561V group. Hence, co-expression of wild-type and A561V mutant KCNH2 in 293 cells activated the UPR process, which led to the inhibition of protein translation and synthesis, in turn inhibiting the expression of KCNH2. These results provided a theoretical basis for clinical treatment of Long QT syndrome.

Background:Long QT syndrome type 2 (LQT2) is caused by mutations in the KCNH2/human ether-à-go-go-related gene (hERG). Some hERG genetic mutation-associated diseases are alleviated by hERG-specific drug chaperones (glycerol, dimethyl sulfoxide, trimethylamine N-oxide, thapsigargin), delayed rectifier K+ current (IKr) blockers methanesulfonanilide E4031, the antihistamine astemizole, or the prokinetic drug cisapride, and the anti-arrhythmic drug quinidine. Meanwhile, many in vivo and in vitro studies have reported the efficacy of 4-phenylbutyric acid (4-PBA) in diseases with inherited genetic mutations. This study aims to explore potential therapeutic agents for hERG/G572R mutated ion channel.Methods:pcDNA3/hERG [wild type (WT)]-FLAG and pcDNA3/hERG (G572R)-FLAG plasmids were transfected into HEK293 cells. A western blot (WB) experiment was conducted to analyze protein expression. Quantitative real-time polymerase chain reaction (qPCR) was used to analyze the messenger RNA (mRNA) expression levels in the WT/G572R heterozygous HEK293 cell model treated with or without 4-PBA. The interaction between WT/G572R and BIP (GRP78), GRP94, and 3-hydroxy-3-methylglutaryl coenzyme A reductase degradation protein 1 (HRD1) was tested by co-immunoprecipitation (co-IP). To investigate the effect of 4-PBA on the WT/G572R channel current, we used electrophysiological assays (patch-clamp electrophysiological recordings).Results:The results showed that WT/G572R activated the ATF6 pathway in the endoplasmic reticulum stress (ERS), the ERS response markers GRP78, GRP94, and calreticulin (CRT)/calnexin (CNX), and HRD1, which decreased after application of the ERS inhibitor 4-PBA. The results of co-IP confirmed that the ability of hERG interacted with GRP78, GRP94, and HRD1. Moreover, 4-PBA increased the current of WT/G572R and reversed the gating kinetics of the WT/G572R channel.Conclusions:4-PBA corrects hERG channel transport defects by inhibiting excessive ERS and the endoplasmic reticulum-associated degradation (ERAD)-related gene E3 ubiquitin ligase HRD1. Additionally, 4-PBA improved WT/G572R channel current. 4-PBA is expected to be developed as a new treatment method for LQT2.

Aims: The progressive development of myocardial remodeling following myocardial infarction (MI) ultimately leads to heart failure (HF). Inflammation has emerged as a crucial catalyst in cardiac remodeling, and inhibiting inflammatory factors holds promise for improved prognosis. In this study, we elucidate the role of IFI-16 (Interferon Gamma Inducible Protein-16) in inducing inflammation through the inflammasome and interacts with galectin-3 signaling pathway in the post-MI heart.Methods and Results: Through analysis of RNA sequencing data from a mouse model of myocardial remodeling available on the Gene Expression Omnibus (GEO) database, we identified ifi202b (human ifi16) as a highly upregulated gene. Our findings reveal elevated levels of IFI-16 (mouse IFI-204) in AC16 human myocytes under hypoxic conditions and in a mouse model of myocardial infarction. Additionally, we observed concomitant upregulation of proteins associated with the inflammasome and galectin-3 signaling pathway. In vitro and in vivo models using shRNA lentivirus or adeno-associated virus (AAVs) to knock down IFI-16/IFI-204 demonstrated reduced activation of the inflammasome and galectin-3 signaling pathway, leading to alleviated remodeling and improved cardiac function during the post-MI procedure. Furthermore, quantitative PCR analysis revealed a decrease in the release of mitochondrial DNA (mtDNA), which correlated with the decline in IFI-16 expression, besides immunofluorescence staining demonstrated significant colocalization between IFI-16 protein and mitochondria.Conclusion: Our findings indicate that IFI-16 plays a pivotal role as a pro-inflammatory factor during the process of post-MI myocardial remodeling. Targeting IFI-16 inhibition may thus present a promising therapeutic strategy for managing myocardial remodeling.Funding: This work was supported by National Natural Science Foundation of China (8200150472); Guangzhou Basic and Applied Basic Research Foundation (2023A04J0448); Post-doctoral research project of Guangdong Provincial Hospital of Chinese Medicine (No. 10187).Declaration of Interest: The authors have declared that no competing interest exists.Ethical Approval: The experimental procedures conducted in this study adhered to the guidelines outlined in the NIH Guide for the Care and Use of Laboratory Animals. Ethical approval for the study was obtained from the Research Institute of the Animal Protection and Use Committee of Guangdong Provincial Hospital of Chinese Medicine [SCXK(Yue) 2021045].

目的 探讨坎地沙坦改善血管紧张素Ⅱ导致的内皮损伤的机制.方法 (1)先用血管紧张素Ⅱ(AngⅡ)干预培养人脐静脉内皮细胞(HUVEC)构建氧化应激细胞株,再用氯沙坦或坎地沙坦对氧化应激的细胞株进行干预,最后用WB法检测eNOS、P-eNOS内皮功能蛋白,P38/P-P38、NF-KB/P-NF-KB炎症通路蛋白及血管紧张素转化酶2(ACE2)蛋白的表达情况.(2)先用AngⅡ和坎地沙坦干预HUVEC细胞构建模型细胞株,再用ACE2的小干扰RNA(siRNA)和ACE2激动剂(DIZE)干预模型细胞株,最后用WB法检测ACE2蛋白,P38/NF-KB通路中P-P38和P-NF-KB蛋白的表达情况.结果 (1)氯沙坦或坎地沙坦干预后,坎地沙坦组和氯沙坦组的P-eNOS蛋白较AngⅡ组的表达含量上升;坎地沙坦组和氯沙坦组P-P38及P-NF-KB的蛋白表达含量较AngⅡ干预组下降,且坎地沙坦组的下降趋势更加明显.(2)予DIZE干预后,HUVECs的ACE2蛋白表达升高,P-NF-KB蛋白的表达下降;予ACE2小干扰RNA干预后,HUVECs的ACE2表达下降;P-NF-KB蛋白的表达上升.结论 坎地沙坦除了可以通过拮抗AngⅡ的受体通路阻断P38/NF-KB炎症通路表达保护内皮细胞功能之外,还可以通过促进内皮细胞分泌ACE2抑制NF-KB炎症通路从而起到保护内皮细胞的作用.

Long QT syndrome type 2 is caused by a mutation in the human-ether-a-go-go-related gene (HERG) gene encoding the rapidly activating delayed rectifier K-current. HERG is a key cell membrane glycoprotein; however, whether the maturation process of HERG protein involves key molecules derived from the calnexin (CNX)/calreticulin (CRT) cycle and how these molecules work remains unknown. Using western blotting, the present study screened the key molecules CNX/CRT/endoplasmic reticulum protein 57 (ERP57) involved in this cycle, and it was revealed that the protein expression levels of CNX/CRT/ERP57 in wild-type (WT)/A561V cells were increased compared with those in WT cells (n=3; P<0.05). Additionally, a co-immunoprecipitation experiment was used to reveal that the ability of CNX/ERP57/CRT to interact with HERG was significantly increased in A561V and WT/A561V cells (n=3; P<0.05). A plasmid lacking the bb′ domain of ERP57 was constructed and it was demonstrated that the key site of ERP57 binding to CRT and immature HERG protein is the bb′ domain. The whole-cell patch-clamp technique detected that the tail current density increased by 46% following overexpression of CRT and by 53% following overexpression of ERP57 in WT/A561V cells. Overexpression of CRT and ERP57 could increased HERG protein levels on the membrane detected by confocal imaging. Furthermore, overexpression of ERP57 and CRT proteins could restore the HERG-A561V mutant protein trafficking process and rescue the dominant-negative suppression of WT. Overall, ERP57/CRT served a crucial role in the HERG-A561V mutant protein trafficking deficiency and degradation process.

1 Targeted inhibition of KDM6 histone demethylases eradicates tumor-initiating cells via enhancer reprogramming in colorectal cancer Junbao Zhang, Ying Ying, Meiqi Li, Maolin Wang, Xiaoyan Huang, Min Jia, Junhui Zeng, Canjie Ma, Yixiang Zhang, Chen Li, Xiaomei Wang, Xing-sheng Shu 1Department of Physiology, School of Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China, 2Department of Urology, The Second Affiliated Hospital of Jinan University, Shenzhen People’s Hospital, Shenzhen 518020, China; 3Department of Oncology, Peking University Shenzhen Hospital, Shenzhen Key Laboratory of Gastrointestinal Cancer Translational Research, Cancer Institute of Shenzhen-PKU-HKUST Medical Center, Shenzhen 518036, China

BackgroundCoronary heart disease (CHD) is primarily caused by atherosclerosis of coronary arteries. It is largely an inflammatory disease of the vascular wall. The inflammation is related to DNA methylation. Angiopoietin‐like protein 2 (ANGPTL2) has various functions in several chronic inflammatory diseases. Macrophage‐derived ANGPTL2 was reported to accelerate CHD development. It is reported that DNA hypomethylation in the promoter region of ANGPTL2 gene was associated with acute coronary syndrome (ACS), a type of CHD. Our objective was to explore the correlation between promoter methylation of the ANGPTL2 gene and CHD, and to investigate the association between methylation status and clinical characteristics of CHD patients.MethodsFirstly, we collected 122 CHD patients and 58 non‐CHD participants from Han Chinese population and purified the peripheral blood DNA. The purified DNA was subjected to bisulfite modification. After bisulfite conversion, the target DNA locus was amplified using polymerase chain reaction (PCR), and the PCR products were measured by pyrosequencing. Finally, the methylation level was calculated according to the sequencing result, and the data were analyzed using xx software.ResultsCHD patients had a relatively lower methylation levels (P50: 7.67% [P25: 6.22%, P75: 10.43%]) in the ANGPTL2 promoter region than did controls (P50: 8.25% [P25: 5.46%, P75: 17.98%], P = 0.001), indicating an association between ANGPTL2 promoter methylation and CHD (OR: 0.890; 95% CI, 0.832‐0.953; adjusted P = 0.001). A breakdown analysis by gender showed that ANGPTL2 promoter methylation was associated with CHD in females (adjusted P = 0.002) but not in males (adjusted P = 0.404). We found no correlation between gene methylation and other clinical characteristics.ConclusionsThe present work provides evidence to support an association between ANGPTL2 promoter DNA methylation status and the risk profile of CHD in females. Our data indicated that in females, promoter DNA hypomethylation of the ANGPTL2 gene is associated with an increased risk of CHD.

Objective To investigate whether RNA (miR-148a) can target DNA transferase 1 (DNMT1) in macrophages,thereby regulating the methylation level of the ATP binding cassette transporter G1 (ABCG1).Methods Bioinformatics analysis was used to predict if DNA methyltransferases 1 (DNMT1) is the target gene of miR-148a.Dual-luciferase reporter assay was used to test whether miR-148a can directly target on the 3'UTR of DNMT1 in HeLa cell.In order to identify if miR-148a could inhibit DNMT1 expression,vctor cloned with the DNMT1 (3'UTR) was co-transfected with miR-148a mimic into THP-1 derived macrophages.Real-time PCR (RT-PCR) was used to test the expression of DNMT1 mRNA and Western Blot was used to analyze the expression of DNMT1 protein.Pyrophosphate assay was used to analyze ABCG1 methylation level from THP-1 derived macrophages after transfecting miR-148a mimic.Results miR-148.a can significantly decreased the luciferase activity of pmirGLO-DNMT3A.miR-148a could not reduce the mRNA level of DNMT1 in THP-1 derived macrophages.miR-148a could inhibit the expression of DNMT1 protein;with all the differences statistically significant(P ＜0.05).The pyrophosphate assay results showed that miR-148a could not down-regulate ABCG1 methylation level in THP-1 derived macrophages.Conclusion miR-148a can directly target on the 3'UTR of DNMT1 and inhibits DNMT1 protein expression,however can not interfere ABCG1 methylation level in macrophage.

miRNAs Regulate hERG. Background: The human ether-a-go-go-related gene (hERG) is the major molecular component of the rapidly activating delayed rectifier K+ current (I-kr). Impairment of hERG function is believed to be amechanism causing long-QT syndromes (LQTS). Growing evidences have shown that microRNAs (miRNAs) are involved in functional modulation of the hERG pathway. The purpose of this study was to screen and validate miRNAs that regulate the hERG pathway. The miRNAs identified in this study will provide new tools to assess the mechanism of LQTS.Methods: Six miRNAs were selected by algorithm predictions based on potential interaction with hERG. The effects of each miRNA on hERG were assessed by use of the Dual-Luciferase Reporter assay system, qRT-PCR, Western blotting, and confocal fluorescence microscopy. Furthermore, whole-cell patch clamp technique was used to validate the effect of miR-103a-1 on the electrophysiological characteristic of the I-kr of the hERG protein channel.Results: miR-134, miR-103a-1, miR-143, and miR-3619 significantly downregulated luciferase activity (P < 0.05) in a reporter test system. These 4 miRNAs significantly suppressed expression of hERG mRNA and protein in U2OS cells (P < 0.05). Corresponding AMOs rescued expression of hERG mRNA and protein. Confocal microscopy showed that all 4 miRNAs reduced the expression of both immature and mature hERG protein. miR-103a-1 decreased the maximum current and tail current amplitudes of hERG channel.Conclusions: Expression and functions of hERG are regulated by specific miRNAs.

Coronary artery disease (CAD) is the leading cause of human morbidity and mortality worldwide. Innovative diagnostic biomarkers are a pressing need for this disease. miRNAs profiling is an innovative method of identifying biomarkers for many diseases and could be proven as a powerful tool in the diagnosis and treatment of CAD. We performed miRNA microarray analysis from the plasma of three CAD patients and three healthy controls. Subsequently, we performed quantitative real-time PCR (qRT-PCR) analysis of miRNA expression in plasma of another 67 CAD patients and 67 healthy controls. We identified two miRNAs (miR-206 and miR-574-5p) that were significantly up-regulated in CAD patients as compared with healthy controls (P<0.05). The receiver operating characteristic (ROC) curves indicated these two miRNAs had great potential to provide sensitive and specific diagnostic value for CAD.

目的 分别构建先天性长QT综合征(LQTS)相关HERG基因和E637K突变基因与红色和绿色荧光蛋白基因的融合表达载体pmCherry-WT-hERG和pEGFP-E637K-hERG,观察其在细胞内的表达和定位情况.方法 将克隆在pcDNA3上的HERG和E637K片段分别亚克隆到红色荧光蛋白载体pmCherry-C2和绿色荧光蛋白载体pEGFP-C1上,转染HEK293T细胞,24 h后利用western blot技术和荧光显微镜观察重组质粒蛋白表达和细胞内定位情况.结果 重组质粒经酶切、PCR和测序鉴定正确无误.转染pmCherry-WT-hERG野生型质粒的细胞可以检测到135 kD和155 kD 2条蛋白质条带,而转染pEGFP-E637K-hERG突变型质粒的细胞仅显示1条135 kD蛋白质条带,155 kD处条带缺失.融合蛋白发出的红色和绿色荧光表明,突变型蛋白分布于胞浆中,而野生型蛋白主要位于胞膜上.结论 成功构建了pmCherry-WT-hERG和pEGFP-E637K-hERG不同荧光蛋白融合表达载体,并在真核细胞中得到有效表达,为LQTS突变基因的进一步功能研究奠定了基础.

This study compared microRNA (miRNA) expression profiles between rheumatic heart disease (RHD) patients and healthy controls to investigate their differential expression and help elucidate their mechanisms of action. Microarray analysis was used to measure miRNA expression, and a total of 133 miRNAs were shown to be significantly upregulated in RHD patients compared with controls, including miR-1183 and miR-1299. A total of 137 miRNAs, including miR-4423-3p and miR-218-1-3p, were significantly downregulated in RHD patients. Quantitative real-time-PCR confirmed microarray findings for miR-1183 and miR-1299 in both tissue and plasma. Bioinformatic predictions were also made of differentially expressed miRNAs as biomarkers in RHD by databases and GO/pathway analysis. Furthermore, we investigated miR-1183 and miR-1299 expression in RHD patients with secondary pulmonary hypertension (PAH). Our findings identified an important role for miR-1299 as a direct regulator of RHD, while the observed difference in expression of miR-1183 between RHD-PAH patients with high or low pulmonary artery pressure suggests that miR-1183 overexpression may reflect pulmonary artery remodeling. miR-1183 and miR-1299 appear to play distinct roles in RHD pathogenesis accompanied by secondary PAH and could be used as potential biological markers for disease development.

Aim: The aim of this study was to assess whether rs1333049 was associated with coronary heart disease (CHD) in Han Chinese.Methods: This case-control study was involved with 599 CHD patients and 591 non-CHD controls. Meanwhile, a comprehensive meta-analysis was also conducted to establish the contribution of rs1333049 to CHD.Results: Our results showed that rs1333049 increased the risk of CHD by 38% (OR=1.38, 95% CI=1.18-1.62). A breakdown analysis by gender further indicated that rs1333049 increased the risk of CHD in men by 29% (OR=1.29, 95% CI=1.05-1.58) and in women by 64% (OR=1.64, 95% CI=1.25-2.16). A follow-up subgroup analysis by age showed there was a significant association between rs1333049 and CHD in women younger than 65 (≤55 years: p=0.001, 55-65 years: p=0.008) and in men aged between 55 and 65 years (p=0.005). Our meta-analysis was involved with 21 studies (25 stages) among 20969 cases and 34114 controls. Our results showed that rs1333049 led to a significantly increased risk of CHD (OR=1.30, 95% CI=1.21-1.39). Further subgroup analyses by ethnicity showed rs1333049 increased the CHD risk by 30% in Europeans (OR=1.30, 95% CI=1.16-1.47) and 27% in Asians (OR=1.27, 95% CI=1.22-1.33).Conclusions: Our case-control study and meta-analysis suggest that rs1333049 is a useful risk marker of CHD.

The goal of our study is to test the association of IL6R rs7529229 polymorphism with CHD through a case-control study in Han Chinese population and a meta-analysis. Our result showed there is a lack of association between IL6R rs7529229 polymorphism and CHD on both genotype and allele levels in Han Chinese (P > 0.05). However, a meta-analysis among 11678 cases and 12861 controls showed that rs7529229-C allele was significantly associated with a decreased risk of CHD, especially in Europeans (P < 0.0001, odds ratio -0.93, 95% confidential interval -0.89-0.96). Since there is significant difference among different populations, further studies are warranted to test the contribution of rs7529229 to CHD in other ethnic populations.

Endothelial progenitor cells (EPCs) are bone marrow-derived cells that have the propensity to differentiate into mature endothelial cells (ECs). The transplantation of EPCs has been shown to enhance in vivo postnatal neo-vasculogenesis, as well as repair infarcted myocardium. Via the whole-cell patch clamp technique, numerous types of ion channels have been detected in EPCs, including the inward rectifier potassium channel (IKir), Ca2+-activated potassium channel (IKCa), and volume-sensitive chloride channel, but their influence on the differentiation of EPCs has yet to be characterized. The present study was designed to investigate: (1) which ion channels have the most significant impact on the differentiation of EPCs; (2) what role ion channels play in the functional development of EPCs; (3) the mRNA and protein expression levels of related ion channel subunits in EPCs. In our study, EPCs were obtained from the peripheral blood of healthy adults and cultured with endothelial growth factors. When EPCs differentiate into mature ECs, they lose expression of the stem cell/progenitor marker CD133, as analyzed by flow cytometry (0.44±0.20 %). However, treatment with the potassium channel inhibitor, tetraethylammonium (TEA) results in an increase in CD133+ cells (25.50±7.55 %). In a functional experiment, we observed a reduction in the capacity of TEA treated ECs (differentiated from EPCs) to form capillary tubes when seeded in Matrigel. At the mRNA and protein levels, we revealed several K+ subtypes, including KCNN4 for IKCa, KCNNMA1 for BKCa and Kir3.4 for IKir. These results demonstrate for the first time that potassium channels play a significant role in the differentiation of EPCs. Moreover, inhibition of potassium channels may depress the differentiation of EPCs and the significant potassium channel subunits in EPCs appear to be IKCa, BKCa and Kir3.4.

Coronary artery disease (CAD) has become the main cause of mortality worldwide. Lectin galactoside-binding soluble-2 (LGALS2) is involved in the cytokine lymphotoxin-α (LTA) cascade that may influence the progress of CAD. The aim of the present study was to assess the association between the LGALS2 3279C>T (rs7291467) polymorphism and CAD. A total of 562 cases and 572 controls were recruited to examine the association. A systematic meta-analysis was performed to evaluate the contribution of LGALS2 3279C>T polymorphism to the risk of CAD among 12,093 cases and 11,020 controls. There was no significant association found in the present case-control study. However, the meta-analysis showed that LGALS2 3279C>T played a protective role in CAD [P=0.008, odds ratio (OR), 0.90; 95% confidence interval (95% CI), 0.82-0.97] and particularly in the Asian population (P=0.006; OR, 0.82; 95% CI, 0.71-0.94). The present case-control study did not find a significant association between LGALS2 3279C>T and CAD in the Eastern Han Chinese population. However, the meta-analysis indicated that LGALS2 3279C>T played a protective role in CAD, suggesting an ethnic difference in the association of the locus with CAD.

To investigate the association of ABCG1, GALNT2 and HMGCR genes promoter DNA methylation with coronary heart disease (CHD) and explore the interaction between their methylation status and the CHD patients' clinical characteristics in Han Chinese population.Methylation-specific polymerase chain reaction (MSP) technology was used to examine the role of the aberrant gene promoter methylation in CHD in Han Chinese population. A total of 85 CHD patients and 54 participants without CHD confirmed by angiography were recruited. 82.8% of the participants with ABCG1 gene promoter hypermethylation have CHD, while only 17.4% of the participants without hypermethylation have it. The average age of the participants with GALNT2 gene promoter hypermethylation is 62.10 ± 8.21, while that of the participants without hypermethylation is 57.28 ± 9.87; in the former group, 75.4% of the participants have CHD, compared to only 50% in the latter group. As for the HMGCR gene, the average age of the participants with promoter hypermethylation is 63.24 ± 8.10 and that of the participants without hypermethylation is 57.79 ± 9.55; its promoter hypermethylation is likely to be related to smoking. Our results indicated a significant statistical association of promoter methylation of the ABCG1 gene with increased risk of CHD (OR = 19.966; 95% CI, 7.319-54.468; P*<0.001; P*: adjusted for age, gender, smoking, lipid level, hypertension, and diabetes). Similar results were obtained for that of the GALNT2 gene (OR = 2.978; 95% CI, 1.335-6.646; P* = 0.008), but not of HMGCR gene (OR = 1.388; 95% CI, 0.572-3.371; P* = 0.469).The present work provides evidence to support the association of promoter DNA methylation status with the risk profile of CHD. Our data indicates that promoter DNA hypermethylation of the ABCG1 and GALNT2 genes, but not the HMGCR gene, is associated with an increased risk of CHD. CHD, smoking and aging are likely to be the important factors influencing DNA hypermethylation.